Hayabusa Nozawa

Increase in B-cell-activation factor (BAFF) and IFN-γ productions by tonsillar mononuclear cells stimulated with deoxycytidyl-deoxyguanosine oligodeoxynucleotides (CpG-ODN) in patients with IgA nephropathy

IgA nephropathy (IgAN), the most common form of primary glomerulonephritis, is recognized as a tonsil-related diseases since it often gets worse after and/or during acute tonsillitis and the disease progression is often prevented by tonsillectomy. Although several reports showed an increase in IgA production of tonsillar mononuclear cells (TMCs), its mechanism has not yet been fully clarified. Recently, B-cell-activation factor (BAFF), which stimulates B-cell proliferation and immunoglobulin production, was identified. Unmethylated deoxycytidyl-deoxyguanosine oligodeoxynucleotide (CpG-ODN), which is able to mimic the immunostimulatory activity of microbial DNA, is known to be involved in the production of immunoglobulins and some cytokines. In this study, we focused on roles of BAFF and IFN-gamma in IgA production of TMCs stimulated with CpG-ODN in IgAN patients. Two-color flow cytometric analysis revealed that the intercellular expression of IFN-gamma on the T-cells freshly isolated from tonsils was significantly higher in IgAN patients than in non-IgAN patients (p=0.032). The spontaneous productions of IgA and IFN-gamma of TMCs were significantly higher in IgAN patients than in non-IgAN patients (p=0.023 and p=0.02). Under stimulation with CpG-ODN, the productions of IgA, BAFF and IFN-gamma of TMCs were significantly higher in IgAN patients than in non-IgAN patients (p=0.013, p=0.005 and p=0.039). The IgA production of TMCs stimulated by CpG-ODN was inhibited by the treatment with anti-BAFF antibody and/or anti-IFN-gamma antibody. Under stimulation with IFN-gamma, the BAFF expression on the CD1c cells and the BAFF production of TMCs were significantly higher in IgAN patients than in non-IgAN patients (p=0.004 and p=0.042). These data suggest that hyper-immune response to microbial DNA may be present in IgAN patients and may lead to hyperproduction of BAFF up-regulated by IFN-gamma, resulting in hyperproduction of IgA in IgAN patients.

Up-regulation of CC chemokine receptor 6 on tonsillar T cells and its induction by in vitro stimulation with α-streptococci in patients with pustulosis palmaris et plantaris

Pustulosis palmaris et plantaris (PPP) is a tonsil‐related disease; tonsillectomy is somewhat effective in treating the condition. However, the aetiological association between the tonsils and PPP has not yet been elucidated fully. Recently, some chemokines and chemokine receptors, including CC chemokine receptor (CCR) 4, CCR6 and CX chemokine receptor (CXCR) 3, have been reported to play important roles in the development of psoriasis, a disease related closely to PPP. In this study, we found that CCR6 expression on both tonsillar and peripheral blood T cells was up‐regulated more intensively in PPP patients than in non‐PPP patients (P < 0·001 for both), but CCR4 and CXCR3 expressions were not. In vitro stimulation with α‐streptococcal antigen enhanced CCR6 expression significantly on tonsillar T cells in PPP patients (P < 0·05), but this was not observed in non‐PPP patients. The chemotactic response of tonsillar T cells to the CCR6 ligand CC chemokine ligand (CCL) 20 was significantly higher in PPP patients than in non‐PPP patients (P < 0·05). The percentage of CCR6‐positive peripheral blood T cells decreased after tonsillectomy in PPP patients (P < 0·01); this decrease correlated with an improvement of skin lesions (P < 0·05, r = −0·63). The numbers of CCR6‐positive cells and the expression of CCL20 were increased significantly in pathological lesions compared with non‐pathological lesions in PPP skin (P < 0·01, P < 0·05 respectively). These results suggest that a novel immune response to α‐streptococci may enhance CCR6 expression on T cells in tonsils and that CCR6‐positive T cells may move to peripheral blood circulation, resulting in recruitment to target skin lesions expressing CCL20 in PPP patients. This may be one of the key roles in pathogenesis of the tonsil‐related disease PPP.

Selective expansion of T cell receptor (TCR) V beta 6 in tonsillar and peripheral blood T cells and its induction by in vitro stimulation with Haemophilus parainfluenzae in patients with IgA nephropathy

IgA nephropathy (IgAN), the most common form of primary glomerulonephritis, is recognized as a disease that often becomes worse during acute tonsillitis. Although many reports have shown that tonsillectomy is an effective treatment for IgAN patients, the immunological evidence has not yet been investigated fully. In this study, we compared the expression of T cell receptor (TCR) V beta families in tonsillar T cells of IgAN patients to those of non‐IgAN patients. The reverse transcription–polymerase chain reaction (RT–PCR) and flow cytometric analyses showed that the TCR V beta 6 was used more frequently in tonsillar T cells of IgAN patients than in those of non‐IgAN patients (P < 0·01 each). Similarly, the proportions of TCR V beta 6‐positive cells in peripheral blood T cells were significantly higher in IgAN patients than in non‐IgAN patients (P < 0·05). After tonsillectomy, the proportions decreased in IgAN patients (P < 0·05), but did not in non‐IgAN patients. Furthermore, in vitro stimulation with Haemophilus parainfluenzae antigen, which is reported to deposit in the glomerular mesangium of IgAN, enhanced expression of TCR V beta 6 in tonsillar T cells from both IgAN and non‐IgAN patients. These results suggest that TCR V beta 6‐positive tonsillar T cells might be activated by H. parainfluenzae, move into the kidney through blood circulation and induce glomerulonephritis.

Tonsil immunity and efficacy of tonsillectomy for tonsillar focal infections

Abstract To clarify the efficacy of tonsillectomy for pustulosis palmaris et plantaris (PPP) and immunologic features of tonsils with PPP, we monitored skin improvement after tonsillectomy and investigated tonsillar T-cell subsets expressed with co-stimulatory factors such as CD28 and cytotoxic T cell-associated lymphocyte antigen 4 (CTLA4) and cutaneous lymphocyte antigen (CLA). In our series of 53 patients with PPP, all patients showed 50% skin improvement by 12 months and showed 80–100% skin improvement by 18 months after tonsillectomy. Immunohistologic analyses showed that CLA+CD3 cells increased in both plantar and tonsillar tissues from PPP. In tonsils with PPP, T cell areas increased and correlated with good efficacy of tonsillectomy. Two-color flow cytometry revealed that CD25+ cells and CLA+ cells increased in tonsillar T cells from PPP. RTPCR analysis revealed that the CTLA4 mRNA level decreased in tonsillar T cells with PPP. In vitro stimulation of α-streptococcal antigens to tonsillar T cells induced expression of CLA in PPP. These data suggest that, in tonsils with PPP, CLA+ T cells may be activated through decreased expression of CTLA4 by the response to α-streptococci and may result in homing of the CLA+ T cells to palmar and plantar skin. Such immunologic features of tonsils with PPP may be one of the evidences for high efficacy of tonsillectomy.

A role of skin homing receptor on tonsillar T-cells from patients with pustulosis palmaris et plantaris

Abstract We investigated the expression of cutaneous lymphocyte antigen (CLA) on tonsillar T-cells from patients with pustulosis palmaris et plantaris (PPP). Two-color immunohistologic analysis was used for CLA expression in tonsillar tissues. Two-color flow cytometric analysis was performed for CLA expression on tonsillar T-cells at freshly isolated and α-streptococcal antigen stimulus conditions. Enzyme-linked immunosorbent assay (ELISA) was employed for the measurement of IL-6 and TGF-β levels in the culture supernatant fluids of tonsillar lymphocytes stimulated by α-streptococcal antigens. Two-color immunohistologic analysis and two-color flow cytometry revealed that the numbers of CLA-positive tonsillar CD3-cells in tissue and freshly isolated condition were significantly higher in patients with PPP than those in patients with non-PPP. Stimulation of α-streptococcal antigens from S. mitis and S. salivalius statistically increased CLA-positive CD3-cells in PPP, but did not in non-PPP. The stimulation of S. sanguis and S. salivalius antigen also increased TGF-β production by tonsillar lymphocytes from PPP, but did not from non-PPP. These results suggest that hyper-immune response to α-streptococci may increase CLA expression of tonsillar T-cells through TGF-β production, and it may play a key role in the pathogenesis of PPP.

Mechanisms on the Development of Tonsillar Focal Infections

Pustulosis palmaris et plantaris (PPP), Sterno-costo-claviclar hyperostosis (SCCH), IgA nephropathy, and some other autoimmune diseases have been regarded as tonsillar focal infections since tonsillectomy is quite effective in the treatment of these diseases. In this paper, we reviewed recent clinicopathological evidence on tonsillar focal infections obtained through our experience as well as in the literature. In addition, we summarized experimental results regarding mechanisms of the development and progression of tonsillar focal infections. It has been speculated that abnormal immune responses in the tonsils may cause or worsen the disease. Therefore, tonsillectomy should be recommended for the treatment of tonsillar focal infections.

T-Cell repertoire in the tonsils of patients with IgA nephropathy

It is known that IgA nephropathy (IgAN) often progresses to end-stage renal failure within a period of 20 years. There are many reports suggesting the relationship between the tonsillar autoimmune response and the pathogenesis of IgA nephropathy, however, definitive evidence has not been detected. Recently, studies of T-cell receptor variable (TCR V) region gene usage in human autoimmune disease showed that the TCR V beta repertoire of pathogenic T cells was highly restricted. In this study, we examined expression of TCR V beta families in tonsils from IgAN patients. The study group consisted of 7 IgAN patients and 7 obstructive sleep apnea syndrome (OSAS) patients. Total RNA was extracted from the tonsils of each patients, and expression of each TCR V beta subfamily gene were examined by reverse transcription-polymerase chain reaction (RT-PCR) method. V beta 6 is more frequently used in tonsils of IgAN patients than in those of OSAS patients (p=0.01), and V beta 20 is more frequently used in OSAS patients than IgAN patients (p=0.01). It was already reported that the frequency of V beta 6 subfamily usage is elevated in both peripheral blood lymphocytes (PBL) and renal tissues of IgAN patients. Therefore, our results suggested that the tonsils may be one of the sources of specific T cells related to the pathogenesis of IgAN.

A Case of Anti-phospholipid Syndrome Presenting Unilateral Sensorineural Hearing Loss.

Anti-phospholipid syndrome (APS) was first reported by Harris in 1985 as one of a variety of clinical features associated with episodes of arterial or venous thrombosis. We report a rare case of acute sensorineural hearing loss due to APS. A 40-year-old woman complained of acute sensorineural hearing loss. She had been diagnosed with APS because of positivity for serum anti-phospholipid antibody and 3 episodes of natural abortion. The hearing loss was improved by treatment with predonisolone, but after 4 months, hearing loss recurred. Since she refused steroid therapy and anticoagulation therapy, continuous administration of aspirin was performed. Six months later, her hearing had recovered. If a patient presents with sudden sensorineural hearing loss, it is important to suspect APS and inquire into histories such as thrombosis and recurrent abortion.

A study of costimulatory factors in tonsils of pustulosis palmaris et plantaris

Abstract Pustulosis palmaris et plantaris (PPP) is known to be closely related to tonsillar focal infections and tonsillectomy is quite effective. Hyper-immune response to alpha streptococci in patients with PPP was reported. On the other hand, costimulatory factors were studied on autoimmune disease. CD28 and cytotoxic T lymphocyte-associated 4 (CTLA4) participate in these factors. CD28 take part in T-cell activation, while CTLA4 take part in T-cell suppression. Therefore, we investigated CD28 and CTLA4 mRNA levels by reverse transcription-polymerase chain reaction (RT-PCR) in tonsillar tissues and CD3 positive lymphocytes from patients with PPP, recurrent tonsillitis (RT), and obstructive sleep apnea syndrome (OSAS). We also investigated the expression levels of CD28 and CTLA4 by flow cytometry analysis in tonsillar mononuclear cells stimulated with alpha streptococci in these patients. It was revealed that the CTLA4 mRNA in tonsillar CD3 positive lymphocytes from PPP patients was expressed at lower level than non-PPP patients. Then, the expression levels of CTLA4, comparison before and after stimulation, were lower level in PPP patients than non-PPP patients. These results suggest that the lower expression level of CTLA4 in PPP may cause hyper-immune response to alpha streptococci and abnormal regulation of T-cell activation.

T-cell activation in tonsils of patients with pustulosis palmaris et plantaris

Abstract Pustulosis palmaris et plantaris (PPP) is a well-known skin disease closely related to tonsillar focal infections, and tonsillectomy is very effective for treatment of this condition. However, etiology of PPP was unclear. In this study, we investigated the characteristics of tonsils from PPP patients by clinical, immunohistochemical, immunological, and molecular–biological approach, and considered the etiology of PPP. For 47 Japanese patients with PPP who have tonsillectomy in Asahikawa Medical College, the skin lesion of PPP was improved in 87% of PPP patients 12 months after tonsillectomy. In quantitative immunohistologic analysis by measurement of T-cell nodule areas on tonsillar sections from those patients, there was positive correlation between the enlargement and improvement of the skin lesion. These results suggested that the T-cell nodule expansion might be an important clue towards clarifying the pathogenesis of this tonsil related disease. In flow cytometric analysis, CD3+, CD4+ and CD4+CD25+ cells were significantly elevated in tonsillar lymphocyte from PPP patients. In reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting analyses of TGF-beta, IL-10, and Smad group, Smad7 mRNA and protein in tonsillar CD3+ cells from PPP patients were expressed at higher level than those from obstructive sleep apnea syndrome (OSAS) patients. These results suggested that helper T cells may be frequently activated and proliferating in tonsils of PPP patients, and the activation and proliferation of helper T cells may be due to enhanced inhibition by Smad7 in intracellular signal-transduction of TGF-beta.